Saccharomyces cerevisiae lacking Btn1p modulate vacuolar ATPase activity to regulate pH imbalance in the vacuole.
AuthorsPadilla-López S, Pearce DA
JournalJ Biol Chem
PubMed ID16423829
'The vacuolar H(+)-ATPase (V-ATPase) along with ion channels and transporters maintains vacuolar pH. V-ATPase ATP hydrolysis is coupled with proton transport and establishes an electrochemical gradient between the cytosol and vacuolar lumen for coupled transport of metabolites. Btn1p, the yeast homolog to human CLN3 that is defective in Batten disease, ... More
Yeast V-ATPase complexes containing different isoforms of the 100-kDa a-subunit differ in coupling efficiency and in vivo dissociation.
AuthorsKawasaki-Nishi S, Nishi T, Forgac M
JournalJ Biol Chem
PubMed ID11278748
'The 100 kDa a-subunit of the yeast vacuolar (H(+))-ATPase (V-ATPase) is encoded by two genes, VPH1 and STV1. These genes encode unique isoforms of the a-subunit that have previously been shown to reside in different intracellular compartments in yeast. Vph1p localizes to the central vacuole, whereas Stv1p is present in ... More
Subunit D (Vma8p) of the yeast vacuolar H+-ATPase plays a role in coupling of proton transport and ATP hydrolysis.
AuthorsXu T, Forgac M
JournalJ Biol Chem
PubMed ID10801866
'To investigate the function of subunit D in the vacuolar H(+)-ATPase (V-ATPase) complex, random and site-directed mutagenesis was performed on the VMA8 gene encoding subunit D in yeast. Mutants were selected for the inability to grow at pH 7.5 but the ability to grow at pH 5.5. Mutations leading to ... More
Oligodeoxynucleotides covalently linked to intercalating agents: a new family of gene regulatory substances.
AuthorsHéléne C, Montenay-Garestier T, Saison-Behmoaras T, Toulmé JJ, Boidot-Forget M, Cazenave C, Asseline U, Lancelot G, Maurizot JC, Toulmé F
JournalBiochem Soc Trans
PubMed ID3709940
Interactions between the F1 and F0 parts in the Escherichia coli ATP synthase. Associations involving the loop region of C subunits.
AuthorsWatts SD, Capaldi RA
JournalJ Biol Chem
PubMed ID9182524
'The N-ethylmaleimide reactivity of c subunits in Escherichia coli F1F0 ATP synthase (ECF1F0) isolated from five mutants, each with a cysteine at a different position in the polar loop region (positions 39, 40, 42, 43, and 44), has been investigated. The maleimide was found to react with Cys placed at ... More
Acridine dimers: influence of the intercalating ring and of the linking-chain nature on the equilibrium and kinetic DNA-binding parameters.
AuthorsMarkovits J, Garbay-Jaureguiberry C, Roques BP, Le Pecq JB
JournalEur J Biochem
PubMed ID2924770
'The rigidity of the linking chain of bifunctional intercalators in the ditercalinium series was shown to be critical for antitumor activity. In order to study the influence of the rigidity of the linking chain on the DNA-binding properties of DNA bifunctional intercalators, fluorescent 9-aminoacridine and 2-methoxy-6-chloro-9-aminoacridine analogues with chains of ... More
Activity and in vitro reassembly of the coated vesicle (H+)-ATPase requires the 50-kDa subunit of the clathrin assembly complex AP-2.
AuthorsLiu Q, Feng Y, Forgac M
JournalJ Biol Chem
PubMed ID7989329
'We have previously shown that the 50-kDa subunit of the clathrin assembly complex AP-2 (AP50) stoichiometrically binds to and is immunoprecipitated with the vacuolar (H+)-ATPase (V-ATPase) from clathrin-coated vesicles (Myers, M., and Forgac, M. (1993) J. Biol. Chem. 268, 9184-9186). We now report that treatment of stripped coated vesicles with ... More
Site-directed mutagenesis of the yeast V-ATPase B subunit (Vma2p).
AuthorsLiu Q, Kane PM, Newman PR, Forgac M
JournalJ Biol Chem
PubMed ID8567653
'The B subunit of the vacuolar (H+)-ATPase (V-ATPase) has previously been shown to participate in nucleotide binding and to possess significant sequence homology with the alpha subunit of the mitochondrial F-ATPase, which forms the major portion of the noncatalytic nucleotide binding sites and contributes several residues to the catalytic sites ... More
Formation of the b subunit dimer is necessary for interaction with F1-ATPase.
AuthorsSorgen PL, Bubb MR, McCormick KA, Edison AS, Cain BD
JournalBiochemistry
PubMed ID9454582
'In earlier work, we [McCormick, K. A., et al. (1993) J. Biol. Chem. 268, 24683-24691] observed that mutations at Ala-79 of the b subunit affect assembly of F1F0 ATP synthase. Polypeptides modeled on the soluble portion of the b subunit (bsol) with substitutions at the position corresponding to Ala-79 have ... More
Influence of subunit-specific antibodies on the activity of the F0 complex of the ATP synthase of Escherichia coli. I. Effects of subunit b-specific polyclonal antibodies.
AuthorsDeckers-Hebestreit G, Simoni RD, Altendorf K
JournalJ Biol Chem
PubMed ID1376322
'Incubation of F1-stripped everted membrane vesicles with antibodies against subunit b of the ATP synthase from Escherichia coli resulted in an inhibition of the binding of F1 to F0, whereas the proton translocation remained unaffected. Incubation of unstripped everted membrane vesicles with anti-b antibodies resulted in a partial loss of ... More
Function of the COOH-terminal domain of Vph1p in activity and assembly of the yeast V-ATPase.
AuthorsLeng XH, Manolson MF, Forgac M
JournalJ Biol Chem
PubMed ID9506970
'We have previously shown that mutations in buried charged residues in the last two transmembrane helices of Vph1p (the 100-kDa subunit of the yeast V-ATPase) inhibit proton transport and ATPase activity (Leng, X. H., Manolson, M., Liu, Q., and Forgac, M. (1996) J. Biol. Chem. 271, 22487-22493). In this report ... More
Characterization of the membrane domain subunit NuoJ (ND6) of the NADH-quinone oxidoreductase from Escherichia coli by chromosomal DNA manipulation.
AuthorsKao MC, Di Bernardo S, Nakamaru-Ogiso E, Miyoshi H, Matsuno-Yagi A, Yagi T
JournalBiochemistry
PubMed ID15736965
'The ND6 subunit is one of seven mitochondrial DNA-encoded subunits of the proton-translocating NADH-quinone oxidoreductase (complex I). Physiological importance of the ND6 subunit is becoming increasingly apparent because a number of mutations leading to amino acid changes in this subunit have been found to be associated with known mitochondrial diseases. ... More
ATP-dependent spectral response of oxonol VI in an ATP-Pi exchange complex.
AuthorsKiehl R, Hanstein WG
JournalBiochim Biophys Acta
PubMed ID6235853
'Energy transduction in an ATPase complex (complex V) has been studied in two reactions catalyzed by this system, i.e., ATP-dependent spectral shift of oxonol VI, and ATP-Pi exchange activity. Aurovertin alone inhibits 50% of the oxonol shift at 2 microM, and no further inhibition occurs at up to 12 microM. ... More
A triple mutation in the a subunit of the Escherichia coli/Propionigenium modestum F1Fo ATPase hybrid causes a switch from Na+ stimulation to Na+ inhibition.
AuthorsKaim G, Dimroth P
JournalBiochemistry
PubMed ID9521783
'Previously we have shown that the Na+-translocating Escherichia coli (F1-delta)/Propionigenium modestum (Fo+delta) hybrid ATPase acquires a Na+-independent phenotype by the c subunit double mutation F84L, L87V that is reflected by Na+-independent growth of the mutant strain MPC8487 on succinate [Kaim, G., and Dimroth, P. (1995) J. Mol. Biol. 253, 726-738]. ... More
Catalytic activities of mitochondrial ATP synthase in patients with mitochondrial DNA T8993G mutation in the ATPase 6 gene encoding subunit a.
AuthorsBaracca A, Barogi S, Carelli V, Lenaz G, Solaini G
JournalJ Biol Chem
PubMed ID10660580
'We investigated the biochemical phenotype of the mtDNA T8993G point mutation in the ATPase 6 gene, associated with neurogenic muscle weakness, ataxia, and retinitis pigmentosa (NARP), in three patients from two unrelated families. All three carried >80% mutant genome in platelets and were manifesting clinically various degrees of the NARP ... More
On the role of Arg-210 and Glu-219 of subunit a in proton translocation by the Escherichia coli F0F1-ATP synthase.
AuthorsValiyaveetil FI, Fillingame RH
JournalJ Biol Chem
PubMed ID9405480
'A strain of Escherichia coli was constructed which had a complete deletion of the chromosomal uncB gene encoding subunit a of the F0F1-ATP synthase. Gene replacement was facilitated by a selection protocol that utilized the sacB gene of Bacillus subtilis cloned in a kanamycin resistance cartridge (Ried, J. L., and ... More
Evidence for a selective and electroneutral K+/H(+)-exchange in Saccharomyces cerevisiae using plasma membrane vesicles.
AuthorsCamarasa C, Prieto S, Ros R, Salmon JM, Barre P
JournalYeast
PubMed ID8923735
'The existence of a K+/H+ transport system in plasma membrane vesicles from Saccharomyces cerevisiae is demonstrated using fluorimetric monitoring of proton fluxes across vesicles (ACMA fluorescence quenching). Plasma membrane vesicles used for this study were obtained by a purification/reconstitution protocol based on differential and discontinuous sucrose gradient centrifugations followed by ... More
Site-directed mutagenesis of the 100-kDa subunit (Vph1p) of the yeast vacuolar (H+)-ATPase.
AuthorsLeng XH, Manolson MF, Liu Q, Forgac M
JournalJ Biol Chem
PubMed ID8798414
'Vacuolar (H+)-ATPases (V-ATPases) are multisubunit complexes responsible for acidification of intracellular compartments in eukaryotic cells. V-ATPases possess a subunit of approximate molecular mass 100 kDa of unknown function that is composed of an amino-terminal hydrophilic domain and a carboxyl-terminal hydrophobic domain. To test whether the 100-kDa subunit plays a role ... More
Integration of b subunits of unequal lengths into F1F0-ATP synthase.
AuthorsGrabar TB, Cain BD
JournalJ Biol Chem
PubMed ID12842903
'In Escherichia coli the peripheral stalk of F1F0-ATP synthase consists of a parallel dimer of identical b subunits. However, the length of the two b subunits need not be fixed. This led us to ask whether it is possible for two b subunits of unequal length to dimerize in a ... More
Suppressor mutations in F1 subunit epsilon recouple ATP-driven H+ translocation in uncoupled Q42E subunit c mutant of Escherichia coli F1F0 ATP synthase.
AuthorsZhang Y, Oldenburg M, Fillingame RH
JournalJ Biol Chem
PubMed ID7908291
'The Q42E mutation in the polar loop of subunit c of the Escherichia coli F1F0 ATP synthase leads to an uncoupling of H+ translocation through F0 and ATP synthesis/hydrolysis in F1. We have isolated four second-site suppressor mutants in which the coupling defect is corrected. Substitutions for Glu31 in F1 ... More
A novel mechanism for regulation of vacuolar acidification.
AuthorsFeng Y, Forgac M
JournalJ Biol Chem
PubMed ID1400289
'We have recently demonstrated that Cys-254 of the 73-kDa A subunit of the clathrin-coated vesicle (H+)-ATPase is responsible for sensitivity of the enzyme to sulfhydryl reagents (Feng, Y., and Forgac, M. (1992) J. Biol. Chem. 267, 5817-5822). In the present study we observe that for the purified enzyme, disulfide bond ... More
Inhibition of vacuolar H(+)-ATPase by disulfide bond formation between cysteine 254 and cysteine 532 in subunit A.
AuthorsFeng Y, Forgac M
JournalJ Biol Chem
PubMed ID8175752
'We have previously demonstrated that the coated vesicle vacuolar H(+)-ATPase (V-ATPase) can be inactivated by formation of intramolecular disulfide bonds (Feng, Y., and Forgac, M. (1992) J. Biol. Chem. 267, 19769-19772). The disulfide bond responsible for inactivation can be distinguished from other disulfide bonds that form by the fact that ... More
Biochemical properties of vacuolar zinc transport systems of Saccharomyces cerevisiae.
AuthorsMacDiarmid CW, Milanick MA, Eide DJ
JournalJ Biol Chem
PubMed ID12161436
'The yeast vacuole plays an important role in zinc homeostasis by storing zinc for later use under deficient conditions, sequestering excess zinc for its detoxification, and buffering rapid changes in intracellular zinc levels. The mechanisms involved in vacuolar zinc sequestration are only poorly characterized. Here we describe the properties of ... More
Electrochemical potential and ion transport in vesicles of yeast plasma membrane.
AuthorsCalahorra M, Ramírez J, Clemente SM, Peña A
JournalBiochim Biophys Acta
PubMed ID2883994
'Vesicles from yeast plasma membrane were prepared according to Franzusoff and Cirillo [1983) J. Biol. Chem. 258, 3608), with slight modifications. When Mg-ATP was added, this preparation was able to generate a membrane potential, that was sensitive to inhibitors of the yeast H+-ATPase and uncouplers, and could be decreased by ... More
Rat liver plasma membrane Ca2+- or Mg2+-activated ATPase. Evidence for proton movement in reconstituted vesicles.
AuthorsLuu-The V, Goffeau A, Thinès-Sempoux D
JournalBiochim Biophys Acta
PubMed ID2822118
'The Ca2+- or Mg2+-activated ATPase from rat liver plasma membrane was partly purified by treatments with sodium cholate and lysophosphatidylcholine, and by isopycnic centrifugation on sucrose gradients. The ATPase activity had high sensitivity to detergents, poor nucleotide specificity and broad tolerance for divalent cations. It was insensitive to mitochondrial ATPase ... More
Influence of subunit-specific antibodies on the activity of the F0 complex of the ATP synthase of Escherichia coli. II. Effects of subunit c-specific polyclonal antibodies.
AuthorsDeckers-Hebestreit G, Altendorf K
JournalJ Biol Chem
PubMed ID1376323
'After incubation of F1-stripped everted membrane vesicles with antibodies against subunit c of the ATP synthase of Escherichia coli the proton translocation through the open F0 channel was blocked. Rebinding of F1 to those vesicles is affected by the antibody concentration used. In general, the use of F(ab'')2 or Fab ... More
Subunit interactions in the clathrin-coated vesicle vacuolar (H(+))-ATPase complex.
AuthorsXu T, Vasilyeva E, Forgac M
JournalJ Biol Chem
PubMed ID10506135
'The vacuolar (H(+))-ATPases (or V-ATPases) are structurally related to the F(1)F(0) ATP synthases of mitochondria, chloroplasts and bacteria, being composed of a peripheral (V(1)) and an integral (V(0)) domain. To further investigate the arrangement of subunits in the V-ATPase complex, covalent cross-linking has been carried out on the V-ATPase from ... More
Identification of a region involved in the communication between the NADP(H) binding domain and the membrane domain in proton pumping E. coli transhydrogenase.
AuthorsAlthage M, Bizouarn T, Rydström J
JournalBiochemistry
PubMed ID11502193
'The two hydrophilic domains I and III of Escherichia coli transhydrogenase containing the binding sites for NAD(H) and NADP(H), respectively, are located on the cytosolic side of the membrane, whereas the hydrophobic domain II is composed of 13 transmembrane alpha-helices, and is responsible for proton transport. In the present investigation ... More
Purification and reconstitution of the vacuolar H+-ATPases from lemon fruits and epicotyls.
AuthorsMüller ML, Irkens-Kiesecker U, Kramer D, Taiz L
JournalJ Biol Chem
PubMed ID9139735
'The vacuolar H+-ATPases (V-ATPases) of lemon fruits and epicotyls were detergent-solubilized, purified by column chromatography, and reconstituted into artificial proteoliposomes. During purification, a vanadate- and nitrate-sensitive ATPase activity, consisting of partially disassembled V-ATPase complexes, was resolved from the V-ATPase peak. ATPase and H+-transport activities of the purified, reconstituted V-ATPases of ... More
Subunit epsilon of the Escherichia coli ATP synthase: novel insights into structure and function by analysis of thirteen mutant forms.
AuthorsXiong H, Zhang D, Vik SB
JournalBiochemistry
PubMed ID9819235
'Structural models of subunit epsilon of the ATP synthase from Escherichia coli have been determined recently by NMR [Wilkens et al. (1995) Nat. Struct. Biol. 2, 961-967] and by X-ray crystallography [Uhlin et al. (1997) Structure 5, 1219-1230], revealing a two-domain protein. In this study, six new epsilon mutants were ... More
Conformations of duplex structures formed by oligodeoxynucleotides covalently linked to the intercalator 2-methoxy-6-chloro-9-aminoacridine.
AuthorsCieplak P, Rao SN, Hélène C, Montenay-Garestier T, Kollman PA
JournalJ Biomol Struct Dyn
PubMed ID3271480
'A family of covalent complexes between oligonucleotides and derivatives of the intercalating agent 9-amino acridine has been synthesized (Asseline, U., Thuong, N.T. and Helene, C. (1983) C.R.Acad. Sci. (Paris) 297 (III), 369-372) and studied (Lancelot, G., Asseline, U., Thuong, N.T., and Helene, C. (1985) Biochemistry 24, 2521-2529; Lancelot, G., Asseline, ... More
Specific inhibition of mRNA translation by complementary oligonucleotides covalently linked to intercalating agents.
AuthorsToulmé JJ, Krisch HM, Loreau N, Thuong NT, Hélène C
JournalProc Natl Acad Sci U S A
PubMed ID3513172
'Synthetic oligodeoxynucleotides that are covalently linked at their 3'' end to an acridine derivative and are complementary to the repeated sequence UUAAAUUAAAUUAAA adjacent to the ribosome binding site of the gene 32-encoded mRNA from phage T4 have been used to regulate the synthesis of gene 32-encoded protein in vitro. These ... More
Site-directed mutagenesis of tyrosine residues at nicotinamide nucleotide binding sites of Escherichia coli transhydrogenase.
AuthorsOlausson T, Hultman T, Holmberg E, Rydström J, Ahmad S, Glavas NA, Bragg PD
JournalBiochemistry
PubMed ID8241179
'Nicotinamide nucleotide transhydrogenase (E.C.1.6.1.1) from Escherichia coli was investigated with respect to the role of specific conserved tyrosine residues of putative substrate-binding regions. The enzyme from E. coli is made up of two subunits, alpha (510 residues) and beta (462 residues). The corresponding enzyme from bovine mitochondria is a single ... More
Essential aspartate in subunit c of F1F0 ATP synthase. Effect of position 61 substitutions in helix-2 on function of Asp24 in helix-1.
AuthorsZhang Y, Fillingame RH
JournalJ Biol Chem
PubMed ID8106529
'Subunit c of the F1F0 type, H(+)-transporting ATP synthase contains an essential Asp that is thought to function in H+ transport. Subunit c folds as a helical hairpin of two transmembrane helices with the essential Asp centered at residue 61 in transmembrane helix-2. Miller et al. (Miller, M. J., Olderburg, ... More
Protonophoric activity of NADH coenzyme Q reductase and ATP synthase in coupled submitochondrial particles from horse platelets.
AuthorsBaracca A, Bucchi L, Ghelli A, Lenaz G
JournalBiochem Biophys Res Commun
PubMed ID9207178
'A method to prepare coupled submitochondrial particles from horse platelets is described. The method allowed us to study the protonophoric activities of both complex I and complex V following the fluorescence quenching of the monoamine 9-amino-6-chloro-2 methoxyacridine (ACMA), a probe highly sensitive to the generation of a transmembrane delta pH. ... More
The epsilon subunit of the F(1)F(0) complex of Escherichia coli. cross-linking studies show the same structure in situ as when isolated.
AuthorsSchulenberg B, Capaldi RA
JournalJ Biol Chem
PubMed ID10497194
'Four double mutants in the epsilon subunit were generated, each containing two cysteines, which, based on the NMR structure of this subunit, should form internal disulfide bonds. Two of these were designed to generate interdomain cross-links that lock the C-terminal alpha-helical domain against the beta-sandwich (epsilonM49C/A126C and epsilonF61C/V130C). The second ... More
Further evidence for the proton pumping work of tonoplast ATPase from Hevea latex vacuome.
AuthorsMarin B, Blasco F
JournalBiochem Biophys Res Commun
PubMed ID6212055
Active proton uptake in lipid vesicles reconstituted with the purified yeast plasma membrane ATPase. Fluorescence quenching of 9-amino-6-chloro-2-methoxyacridine.
AuthorsDufour JP, Goffeau A, Tsong TY
JournalJ Biol Chem
PubMed ID6213606
Energy-dependent changes of the electrokinetic properties of chloroplasts.
AuthorsSchapendonk AH, Hemrika-Wagner AM, Theuvenet AP, Sang HW, Vredenberg WJ, Kraayenhof R
JournalBiochemistry
PubMed ID7378383
Inhibition of the energy-linked fluorescence response of quinacrine with local anesthetics.
AuthorsMueller DM, Lee CP
JournalFEBS Lett
PubMed ID7067822
Mechanisms of chromosome banding. XI. The ability of various acridine derivatives to cause Q-banding.
AuthorsComings DE, Limon J, Ledochowski A, Tsou KC
JournalExp Cell Res
PubMed ID720423
Fluorescent indicators for intracellular pH.
AuthorsHan J, Burgess K,
JournalChem Rev
PubMed ID19831417
This review is about intracellular pH sensors, includingsmall fluorescent organic molecules, nanoparticles, andfluorescent proteins, e.g., GFP. It focuses on their preparations, photophysical properties, and advantages/disadvantagesfor intracellular pH measurements. The discussion is limitedto fluorescent indicators that have been applied to measureintracellular pH values since 1980. ... More
Functional reconstitution of a proton-translocating system responsive to fusicoccin.
AuthorsAducci P, Ballio A, Blein JP, Fullone MR, Rossignol M, Scalla R
JournalProc Natl Acad Sci U S A
PubMed ID2903497
Crude fusicoccin binding proteins and a partially purified plasma membrane H+-transporting ATPase (EC 3.6.1.34), both solubilized from maize tissues, were simultaneously inserted into liposomes by the freeze-thaw method. ATP-driven intravesicular acidification in the proteoliposomes, measured by the fluorescence quenching of the dye 9-amino-6-chloro-2-methoxyacridine, markedly increased upon addition of fusicoccin to ... More
Assembly of a functional F0 of the proton-translocating ATPase of Escherichia coli.
AuthorsKlionsky DJ, Brusilow WS, Simoni RD
JournalJ Biol Chem
PubMed ID6309770
We have investigated both structural and functional assembly of the F0 portion of the Escherichia coli proton-translocating ATPase in vivo. Fractionation of E. coli minicells containing plasmids which code for parts of the unc operon shows that each of the F0 peptides a, b, and c insert into the cytoplasmic ... More
Electronic factors and acridine frameshift mutagenicity--a pattern recognition study.
AuthorsHenry DR, Lavine BK, Jurs PC
JournalMutat Res
PubMed ID3302688
Using the ADAPT and CHEMLAB-II systems for structure-activity analysis, computer-calculated electronic properties of molecules were used to derive structure-activity relationships for predicting the mutagenicity of a set of substituted acridines in strain TA1537 of the Ames Salmonella assay. A collection of 40 acridines, with a variety of substituents, was examined. ... More
Uptake of fluorescent dyes associated with the functional expression of the cystic fibrosis transmembrane conductance regulator in epithelial cells.
AuthorsWersto RP, Rosenthal ER, Crystal RG, Spring KR
JournalProc Natl Acad Sci U S A
PubMed ID8577734
Specific mutations in the cystic fibrosis transmembrane conductance regulator (CFTR), the most common autosomal recessive fatal genetic disease of Caucasians, result in the loss of epithelial cell adenosine 3',5'-cyclic-monophosphate (cAMP)-stimulated Cl- conductance. We show that the influx of a fluorescent dye, dihydrorhodamine 6G (dR6G), is increased in cells expressing human ... More
Both ATP and the electrochemical potential are required for optimal assembly of pro-OmpA into Escherichia coli inner membrane vesicles.
AuthorsGeller BL, Movva NR, Wickner W
JournalProc Natl Acad Sci U S A
PubMed ID2872675
Pro-OmpA is processed to OmpA by isolated inverted plasma membrane vesicles from Escherichia coli. In the presence of ATP and a membrane potential, 58% (+/- 13%) of the OmpA is sequestered in the vesicles. We sought to determine which of these two metabolic energy sources is used for protein translocation. ... More
Deletions in hydrophilic domains of subunit a from the Escherichia coli F1F0-ATP synthase interfere with membrane insertion or F0 assembly.
AuthorsLewis MJ, Simoni RD
JournalJ Biol Chem
PubMed ID1531341
The a subunit is a membrane component of the F1F0-ATP synthase from Escherichia coli. Regions of a which appear important for membrane insertion or F0 assembly have been identified by analysis of both deletion mutants and fusion proteins which link the mutant a subunits to alkaline phosphatase. This analysis suggests ... More
Some properties of membrane-bound, solubilized and reconstituted into liposomes H+-ATPase of vacuoles of Saccharomyces carlsbergensis.
AuthorsLichko LP, Okorokov LA
JournalFEBS Lett
PubMed ID6147272
Vacuoles of yeast grown in peptone medium possessed high ATPase activity (up to 1 mumol X mg protein-1 X min-1). Membrane-bound and solubilized ATPase activities were insensitive to vanadate and azide, but were inhibited by NO-3 . K+ and cyclic AMP stimulated both membrane-bound and solubilized ATPase activities. Dio-9 activated ... More
Effects of Escherichia coli secB mutations on pre-maltose binding protein conformation and export kinetics.
AuthorsKumamoto CA, Gannon PM
JournalJ Biol Chem
PubMed ID3042772
Mutations affecting the secB gene of Escherichia coli cause a defect in protein export. This report presents the demonstration that the secB mutations caused a defect in co-translational processing of maltose binding protein (MBP). A significant amount of post-translational processing of pre-MBP occurred within 1 min after termination of pulse ... More
A dual role for phosphatidylglycerol in protein translocation across the Escherichia coli inner membrane.
AuthorsKusters R, Breukink E, Gallusser A, Kuhn A, de Kruijff B
JournalJ Biol Chem
PubMed ID8288623
The involvement of phosphatidylglycerol in the SecA-independent translocation of M13 procoat in Escherichia coli was demonstrated. Processing of procoat to mature coat protein was retarded when the level of phosphatidylglycerol was reduced. In vitro translocation experiments using inner membrane vesicles isolated from a strain with inducible synthesis of phosphatidylglycerol, showed ... More
Effect of dimethoate and chlorfenvinphos on plasma membrane integrity of Synechocystis sp. PCC 6803.
AuthorsMohapatra PK, Schiewer U
JournalEcotoxicol Environ Saf
PubMed ID9799578
The organophosphorus (OP) insecticides dimethoate and chlorfenvinphos, at all selected concentrations (0-500 micromol liter-1), reduced the rate of accumulation of uranine and enhanced fluorescence quenching of 9-amino-6-chloro-2-methoxyacridine (ACMA). There was a significant nonlinear negative correlation between insecticide concentrations and uranine accumulation. The reduction in amount of uranine trapped inside the ... More
Insertion scanning mutagenesis of subunit a of the F1F0 ATP synthase near His245 and implications on gating of the proton channel.
AuthorsVik SB, Patterson AR, Antonio BJ
JournalJ Biol Chem
PubMed ID9632681
Subunit a of the E. coli F1F0 ATP synthase was probed by insertion scanning mutagenesis in a region between residues Glu219 and His245. A series of single amino acid insertions, of both alanine and aspartic acid, were constructed after the following residues: 225, 229, 233, 238, 243, and 245. The ... More
Functional expression of plant plasma membrane H(+)-ATPase in yeast endoplasmic reticulum.
AuthorsVillalba JM, Palmgren MG, Berberián GE, Ferguson C, Serrano R
JournalJ Biol Chem
PubMed ID1534807
Recombinant plant plasma membrane H(+)-ATPase has been produced in a yeast expression system comprising a multicopy plasmid and the strong promoter of the yeast PMA1 gene. Western blotting with a specific monoclonal antibody showed that the plant ATPase is one of the major membrane proteins made by the transformed cells, ... More
A Cl(-)-translocating adenosinetriphosphatase in Acetabularia acetabulum. 2. Reconstitution of the enzyme into liposomes and effect of net charges of liposomes on chloride permeability and reconstitution.
AuthorsIkeda M, Oesterhelt D
JournalBiochemistry
PubMed ID2139343
The Mono Q-III fraction, a Mg2(+)-ATPase, isolated from Acetabularia acetabulum was reconstituted into liposomes of various net charges prepared by the reversed-phase method and tested for a Cl(-)-translocating activity. The liposomes from a mixture of egg lecithin, dicetyl phosphate, and cholesterol (63:18:9 mole ratio, negative liposomes) and from a mixture ... More
Second-site suppressor mutations at glycine 218 and histidine 245 in the alpha subunit of F1F0 ATP synthase in Escherichia coli.
AuthorsHartzog PE, Cain BD
JournalJ Biol Chem
PubMed ID7798232
The alpha-like subunits of F1F0 ATP synthases share primary structural homology in two segments near their carboxyl termini. However, the amino acids at the functionally important positions occupied by alpha Gly-218 and alpha His-245 in Escherichia coli vary depending upon organism and organelle. The alpha G218-->D,H245-G and alpha G218-->K,H245-->G double ... More
Interaction between Glu-219 and His-245 within the a subunit of F1F0-ATPase in Escherichia coli.
AuthorsCain BD, Simoni RD
JournalJ Biol Chem
PubMed ID2896197
Oligonucleotide-directed mutagenesis was used to generate mutations in the a subunit gene (uncB) altering the glutamic acid 219 and the histidine 245 codons. Substitutions of aspartic acid, glutamine, histidine, and leucine for glutamic acid at position 219 neither alter the hydrolytic activity of membrane-bound F1 nor the association of F1 ... More
Transmembrane topography of subunit a in the Escherichia coli F1F0 ATP synthase.
AuthorsValiyaveetil FI, Fillingame RH
JournalJ Biol Chem
PubMed ID9632683
Subunit a is the least understood of the three subunits that compose the F0 sector in the Escherichia coli F0F1 ATP synthase. In this study, we have substituted Cys into predicted extramembranous loops of the protein and used chemical modification to obtain topographical information on the folding of subunit a. ... More
Deletions in the second stalk of F1F0-ATP synthase in Escherichia coli.
AuthorsSorgen PL, Caviston TL, Perry RC, Cain BD
JournalJ Biol Chem
PubMed ID9774398
In Escherichia coli F1F0-ATP synthase, the two b subunits form the second stalk spanning the distance between the membrane F0 sector and the bulk of F1. Current models predict that the stator should be relatively rigid and engaged in contact with F1 at fixed points. To test this hypothesis, we ... More
The acridine ring selectively intercalated into a DNA helix at various types of abasic sites: double strand formation and photophysical properties.
AuthorsFukui K, Tanaka K
JournalNucleic Acids Res
PubMed ID8918798
The interactions between the intercalating agent and the three types of abasic sites: abasic frameshift, apurinic and apyrimidinic, were investigated. 9-amino-6-chloro-2-methoxyacridine (ACMA), whose spectroscopic properties are strongly perturbed by the environment, was selected as the intercalating agent. The optically pure threoninol derived from the reduction of L-threonine was used as ... More
The proton channel of the energy-transducing nicotinamide nucleotide transhydrogenase of Escherichia coli.
AuthorsYamaguchi M, Stout CD, Hatefi Y
JournalJ Biol Chem
PubMed ID12087099
The nicotinamide nucleotide transhydrogenases of mitochondria and bacteria are proton pumps that couple direct hydride ion transfer between NAD(H) and NADP(H) bound, respectively, to extramembranous domains I and III to proton translocation by the membrane-intercalated domain II. To delineate the proton channel of the enzyme, 25 conserved and semiconserved prototropic ... More
Mechanistic differences in the energy-linked fluorescence decreases of 9-aminoacridine dyes associated with bovine heart submitochondrial membranes.
AuthorsHuang CS, Kopacz SJ, Lee CP
JournalBiochim Biophys Acta
PubMed ID6824642
(1) The pH dependence of the fluorescence intensities of 9-aminoacridines associated with energized submitochondrial membranes suggests that a mechanism(s) other than protonation of the dye molecules, as is the case with quinacrine, is responsible for the energy-linked fluorescence decreases of 9-aminoacridine and 9-amino-3-chloro-7-methoxyacridine (9-ACMA). (2) That the fluorescence polarization of ... More
Single amino acid insertions probe the alpha subunit of the Escherichia coli F1F0-ATP synthase.
AuthorsWang S, Vik SB
JournalJ Biol Chem
PubMed ID8300644
Single amino acid insertions of alanine or aspartate have been introduced into the alpha subunit of the F1F0-ATP synthase at seven different sites, after residues 187, 193, 198, 202, 212, 217, and 222. These sites span a highly conserved region of the alpha subunit, parts of which are thought to ... More
Characterization of reconstituted ATPase complex proteoliposomes prepared from the thermophilic cyanobacterium Synechococcus 6716.
Authorsvan Walraven HS, Lubberding HJ, Marvin HJ, Kraayenhof R
JournalEur J Biochem
PubMed ID6197302
The preparation and some properties are described of proteoliposomes consisting of the ATPase complex and lipids from the thermophilic cyanobacterium Synechococcus 6716. In the proteoliposomes (about 200 nm in diameter) only a low amount of protein can be incorporated (protein/lipid ratio of 0.01 w/w) and they show very few protein ... More
Photoaffinity labelling of submitochondrial membranes with the 3-azido analogue of 9-amino-3-chloro-7-methoxyacridine.
AuthorsKopacz SJ, Mueller DM, Lee CP
JournalBiochim Biophys Acta
PubMed ID3978094
9-Amino-3-azido-7-methoxyacridine has been synthesized and shown to be a suitable photoaffinity probe for the site(s) of interaction of 9-amino-3-chloro-7-methoxyacridine with submitochondrial membranes. Both the excitation and emission spectra of the azido analogue covalently bound to membranes in the energized state display distinctive differences from the spectra of labelled, non-energized membranes ... More
Catalytic site nucleotide binding and hydrolysis in F1F0-ATP synthase.
AuthorsLöbau S, Weber J, Senior AE
JournalBiochemistry
PubMed ID9692975
F1F0-ATP synthase was purified from Escherichia coli beta Y331W mutant. The beta-Trp-331 provided a specific fluorescent probe of catalytic site nucleotide binding. Physiological (mM) concentration of substrate MgATP filled all three catalytic sites. With MgATP or MgADP the catalytic sites showed marked binding cooperativity and asymmetry, which was dependent on ... More
Lengthening the second stalk of F(1)F(0) ATP synthase in Escherichia coli.
AuthorsSorgen PL, Bubb MR, Cain BD
JournalJ Biol Chem
PubMed ID10593914
In Escherichia coli F(1)F(0) ATP synthase, the two b subunits dimerize forming the peripheral second stalk linking the membrane F(0) sector to F(1). Previously, we have demonstrated that the enzyme could accommodate relatively large deletions in the b subunits while retaining function (Sorgen, P. L., Caviston, T. L., Perry, R. ... More
p-nitrophenylphosphatase activity of plasma membrane H(+)-ATPase from yeast. Implications for the regulation of the catalytic cycle by H+.
AuthorsFerreira-Pereira A, Alves-Ferreira M, de Carvalho-Alves PC
JournalJ Biol Chem
PubMed ID8163511
The H(+)-ATPase from Schizosaccharomyces pombe belongs to the group of transport ATPases which displays two main conformational states, E1 and E2 (P-type ATPase). In this report, we show that, as in the case of other P-type ATPase, the purified enzyme exhibits a p-nitrophenylphosphatase activity which can be completely inhibited by ... More
The proton pumping activity of H(+)-ATPases: an improved fluorescence assay.
AuthorsRottenberg H, Moreno-Sanchez R
JournalBiochim Biophys Acta
PubMed ID8399374
A new method for the estimation of steady-state delta pH, and the rate of acidification, by H(+)-ATPases (and other proton transporters) in inverted membrane vesicles is described. The method is based on a combination of two widely used fluorescent delta pH probes, 9-aminoacridine and 9-amino-6-chloro-2-methoxyacridine. It is demonstrated that 9-amino-6-chloro-2-methoxyacridine ... More
The plant inorganic pyrophosphatase does not transport K+ in vacuole membrane vesicles multilabeled with fluorescent probes for H+, K+, and membrane potential.
AuthorsRos R, Romieu C, Gibrat R, Grignon C
JournalJ Biol Chem
PubMed ID7876200
It has been claimed that the inorganic pyrophosphatase (PPase) of the plant vacuolar membrane transports K+ in addition to H+ in intact vacuoles (Davies, J. M., Poole, R. J., Rea, P. A., and Sanders, D. (1992) Proc. Natl. Acad. Sci. U.S.A. 89, 11701-11705). Since this was not confirmed using the ... More
Mutational analysis of the non-homologous region of subunit A of the yeast V-ATPase.
AuthorsShao E, Nishi T, Kawasaki-Nishi S, Forgac M
JournalJ Biol Chem
PubMed ID12569096
Subunit A is the catalytic nucleotide binding subunit of the vacuolar proton-translocating ATPase (or V-ATPase) and is homologous to subunit beta of the F(1)F(0) ATP synthase (or F-ATPase). Amino acid sequence alignment of these subunits reveals a 90-amino acid insert in subunit A (termed the non-homologous region) that is absent ... More
Cysteine scanning mutagenesis of the noncatalytic nucleotide binding site of the yeast V-ATPase.
AuthorsVasilyeva E, Liu Q, MacLeod KJ, Baleja JD, Forgac M
JournalJ Biol Chem
PubMed ID10617613
To investigate residues involved in the formation of the noncatalytic nucleotide binding sites of the vacuolar proton-translocating adenosine triphosphatase (V-ATPase), cysteine scanning mutagenesis of the VMA2 gene that encodes the B subunit in yeast was performed. Replacement of the single endogenous cysteine residue at position 188 gave rise to a ... More
Antagonism of immunostimulatory CpG-oligodeoxynucleotides by quinacrine, chloroquine, and structurally related compounds.
AuthorsMacfarlane DE, Manzel L
JournalJ Immunol
PubMed ID9570525
Phosphorothioate oligodeoxynucleotides containing CpG (CpG-ODN) activate immune responses. We report that quinacrine, chloroquine, and structurally related compounds completely inhibit the antiapoptotic effect of CpG-ODN on WEHI 231 murine B lymphoma cells and inhibit CpG-ODN-induced secretion of IL-6 by WEHI 231. They also inhibit IL-6 synthesis and thymidine uptake by human ... More
Studies on Na+ and H+ translocation through the Fo part of the Na(+)-translocating F1Fo ATPase from Propionigenium modestum: discovery of a membrane potential dependent step.
AuthorsKluge C, Dimroth P
JournalBiochemistry
PubMed ID1472503
The purified ATPase of Propionigenium modestum (F1Fo) was incorporated into liposomes, and the F1 part was dissociated. The Fo-liposomes catalyzed proton uptake in response to a potassium diffusion potential (inside negative). Proton translocation was abolished by rebinding F1 to the Fo-liposomes or after incubation with the c-subunit-specific inhibitor dicyclohexylcarbodiimide (DCCD). ... More
Interaction of the clathrin-coated vesicle V-ATPase with ADP and sodium azide.
AuthorsVasilyeva E, Forgac M
JournalJ Biol Chem
PubMed ID9726993
The kinetics of adenosine triphosphate (ATP)-dependent proton transport into clathrin-coated vesicles from bovine brain have been studied. We observe that the vacuolar proton-translocating ATPase (V-ATPase) from clathrin-coated vesicles is subject to two different types of inhibition by ADP. The first is competitive inhibition with respect to ATP, with a Ki ... More
Microtubules are involved in glucose-dependent dissociation of the yeast vacuolar [H+]-ATPase in vivo.
AuthorsXu T, Forgac M
JournalJ Biol Chem
PubMed ID11331282
The vacuolar [H(+)]-ATPases (V-ATPases) are composed of a peripheral V(1) domain and a membrane-embedded V(0) domain. Reversible dissociation of the V(1) and V(0) domains has been observed in both yeast and insects and has been suggested to represent a general regulatory mechanism for controlling V-ATPase activity in vivo. In yeast, ... More
Effects of sequential deletions of residues from the N- or C-terminus on the functions of epsilon subunit of the chloroplast ATP synthase.
AuthorsShi XB, Wei JM, Shen YK
JournalBiochemistry
PubMed ID11535058
Ten truncated mutants of chloroplast ATP synthase epsilon subunit from spinach (Spinacia oleracea), which had sequentially lost 1-5 amino acid residues from the N-terminus and 6-10 residues from the C-terminus, were generated by PCR. These mutants were overexpressed in Escherichia coli, reconstituted with soluble and membrane-bound CF(1), and the ATPase ... More
Cross-linking of the delta subunit to one of the three alpha subunits has no effect on functioning, as expected if delta is a part of the stator that links the F1 and F0 parts of the Escherichia coli ATP synthase.
AuthorsOgilvie I, Aggeler R, Capaldi RA
JournalJ Biol Chem
PubMed ID9195980
A mutant of the Escherichia coli F1F0-ATPase has been generated (alphaQ2C) in which the glutamine at position 2 of the alpha subunit has been replaced with a cysteine residue. Cu2+ treatment of ECF1 from this mutant cross-linked an alpha subunit to the delta subunit in high yield. Two different sites ... More
Changing the ion binding specificity of the Escherichia coli H(+)-transporting ATP synthase by directed mutagenesis of subunit c.
AuthorsZhang Y, Fillingame RH
JournalJ Biol Chem
PubMed ID7814424
Most F1F0 type ATP synthases, including that in Escherichia coli, use H+ as the coupling ion for ATP synthesis. However, the structurally related F1F0 ATP synthase in Propionigenium modestum uses Na+ instead. The binding site for Na+ residues in the F0 sector of the P. modestum enzyme. We postulated that ... More
Mechanical inhibition of isolated Vo from V/A-ATPase for proton conductance.
Authors
JournalElife
PubMed ID32639230
Gating and selectivity mechanisms for the lysosomal K+ channel TMEM175.
Authors
JournalElife
PubMed ID32228865
Cryo-EM structures and functional characterization of murine Slc26a9 reveal mechanism of uncoupled chloride transport.
Authors
JournalElife
PubMed ID31339488
Structural and functional properties of a magnesium transporter of the SLC11/NRAMP family.